摘要
目的：利用自发性高血压大鼠(spontaneously hypertensive rat, SHR)模型,观察血管内皮功能障碍标志物与早期肾损害的相关性;同时,在SHR肾损害早期(16周龄始)给予加味天麻钩藤饮进行干预,观察其对SHR血管内皮功能及早期肾损害的作用,探讨效应途径和机制。
方法：购买40只15周龄的同源同系、雄性SHR,适应性喂养一周,称重、标记终身号并随机分组,即模型对照组、加味天麻钩藤饮组、福辛普利组和联合于预组。观察模型对照组SHR 24小时尿蛋白定量与血清一氧化氮(nitric oxide, NO)、血浆(endothelin-1, ET-1)及循环NO/ET-1的相关性；同时,实验各组在高血压肾损害早期(16周龄始)给予相应干预药物,以观察加味天麻钩藤饮、福辛普利及两药联合应用后,对SHR血压、24小时尿蛋白定量、内生肌酐清除率(endogenous creatinine clearance rate, Ccr)、血清NO、血浆ET-1及循环NO/ET-1的影响。
结果：1.模型组SHR24小时尿蛋白排泄量与血清NO呈负相关,Pearson系数为—0.818,P-0.047,P<0.05；与循环NO/ET-1呈负相关,Pearson系数为—0.835,P=0.039,P<0.05；与血清ET-1关系不明显,Pearson系数为0.058,P=0.913,P>0.05。2.加味天麻钩藤饮可以显著降低SHR脉压(pulse pressure, PP),与模型对照组相比,P<0.01；与福辛普利联合应用后,两者降压起协同作用。3.加味天麻钩藤饮可以显著降低血浆ET-1水平,与模型对照组相比,P<0.01；对血清NO水平影响不明显,P>0.05。4.加味天麻钩藤饮与福辛普利联合应用后,24小时尿蛋白定量明显下降,与模型对照组比较,P<0.01；循环NO/ET-1升高,与模型对照组比较,P<0.05。
结论：1.SHR早期肾损害24小时尿蛋白排泄量与血清NO浓度及循环NO/ET-1水平呈负相关,推测高血压早期肾损害与血管内皮功能障碍(endothelial dysfunction, ED)有一定的相关性,尤与血管内皮舒张功能障碍密切相关。2.加味天麻钩藤饮单独应用,即可显著降低脉压,疗效与血管紧张素转化酶抑制剂(angiotensin-converting enzyme inhibitor, ACEI)福辛普利相当。3.加味天麻钩藤饮可以显著降低血浆ET-1水平,改善内皮依赖性收缩功能,推测具有一定的血管内皮保护作用。4.加味天麻钩藤饮与ACEI联合应用后,协同降压、进一步减轻早期肾损害,推测其机制可能与加味天麻钩藤饮的血管内皮保护作用有关。5.加味天麻钩藤饮与ACEI联合应用后,血管内皮功能及肾损害均有一定程度的改善,结合本实验结论1,推测改善血管内皮功能可能是干预高血压早期肾损害的新的有效途径。6.对高血压早期肾损害的防治,强调早期检测敏感指标,并结合辨证,给予加味天麻钩藤饮联合降压药物,进行多靶点干预,能够起到协同增效的作用,更好的保护靶器官功能。
Objective:To observe the relationship between the vascular endothelial dysfunction and early kidney damage by using the model of spontaneously hypertensive rat (SHR); meanwhile, the SHR were fed with the decoction of Jiaweitianmagoutengyin in the early renal damage(16 weeks), to observe its effects on vascular endothelial function and early kidney damage, explore the channels and mechanisms simultaneously.
Methods:40 homogenous 15-week-old male SHR were purchased, adaptive feeding for one week, then weigh, tag the life number and randomly divided into 4 groups, namely, model control group, Jiaweitianmagoutengyin group, Fosinopril group and the joint intervention group. The correlation between 24-hour urine protein excretion and serum NO, plasma ET-1 and circulating NO/ET-1 were observed through the model control group; Meanwhile, the experimental groups, were given corresponding drug-interventions in the early stage of hypertensive renal injury (16 weeks), the changes of blood pressure, 24-hour urine protein quantitative, creatinine clearance rate, serum NO, plasma ET-1 and circulating NO/ET-1 of SHR were observed after using Jiaweitianmagoutengyin decoction, Fosinopril and the combining use of both drugs.
Results:1.In model control group,24-hour urine protein excretion was negatively correlated with plasma ET-1, Pearson coefficient was -0.818, P=0.047, P<0.01; and had a negative correlation with circulating NO/ET-1, too, Pearson coefficient was -0.835, P= 0.039, P<0.05; significant relationship between 24-hour urine protein excretion and serum NO was not found.2.Jiaweitianmagoutengyin decoction can significantly reduce the pulse pressure of SHR, P<0.01, play a synergistic effect of depressurization after combining use of Fosinopril.3.Jiaweitianmagoutengyin decoction can significantly reduce the plasma ET-1 level, P<0.01; no significant effect on serum NO level, P> 0.05.4. After combining use of Jiaweitianmagoutengyin decoction and Fosinopril, the 24-hour urinary protein quantitative decreased significantly, compared with the model control group, P <0.01; circulating NO/ET-1 increased compared with model control group, P<0.05.
Conclusion:1.SHR's 24 hours urinary protein excretion and serum nitric oxide concentration was negatively correlated, there was also a negative correlation between 24 hours urinary protein excretion and the level of circulating NO/ET-1, speculating that there was a certain degree of relevance between the early stage of hypertensive renal damage and vascular endothelial dysfunction.,especially the diastolic dysfunction.2.Independent use of Jiaweitianmagoutengyin decoction can significantly reduce the pulse pressure, has the equal efficacy compared with Fosinopril(ACEI).3. Jiaweitianmagoutengyin decoction has a good protective effect on vascular endothelium, particularly in reducing plasma ET-1 level, mainly to ameliorate the endothelium-dependent contraction.4.Combining use of Jiaweitianmagoutengyin decoction and Fosinopril(ACEI) showed a synergistic effect of depressurization, further ease early kidney damage, speculating its mechanism may be related to the role of vascular endothelial protection of Jiaweitianmagoutengyin decoction. 5.After combining use of Jiaweitianmagoutengyin decoction and Fosinopril(ACEI), vascular endothelial function and renal damage have improved to some extent,considering the first result of this experiment, speculating that improving vascular endothelial function is a new and effective way to intervene early hypertensive renal damage.6.For prevention and treatment of early hypertensive renal damage, we emphasize the early detection of sensitive indicators;combined with syndrome differentiation, joint use of Chinese medicine compound and antihypertensive drugs, multi-target interference,can play a synergistic role for protection of target organs.
引文
[1]王海燕,李晓玫,赵明辉,等.肾脏病学[M].北京：人民卫生出版社,2008,第3版：1662.
[2]卫生部心血管病防治中心,中国心血管病报2005.中国大百科全书出版社,2006：84.
[3]Nissenson AR, Pereira BJ, Collins AJ, et al.Prevalence and characteristics of individuals with chronic kidney disease in a large health maintenance organization[J].Am J Kidney Dis,2001; 37:1177-1183.
[4]Collins AJ, Kasiske B, Herzog C, et al.Excerpts from the United States Renal Data System 2003 Annual Data Report:atlas of end-stage renal disease in the United States[J].Am J Kid Dis,2003; 42:a5-7, S1-S230.
[5]周福德,王梅.北京市血液透析的发展与质量改进[J].中国血液净化杂志,2006；5：117-118.
[6]袁伟杰,崔若兰.对高血压肾损害临床应进一步思考的问题[J].中华肾脏病杂志,2005：21(10)：572-574.
[7]Spencer CG, Martin SC, Felmeden DC, et al. Relationship of homocysteine to markers of platelet and endothelial activation in"high risk" hypertensives:a substudy of the Anglo-Scandinavian Cardiac Outcomes Trial [J].Int J Cardiol,2004; 94(2-3): 293-300.
[8]张会敏,何芳.内皮细胞功能障碍与原发性高血压关系研究进展[J].医学综述,2007：13(2)：114-116.
[9]钟海兰,卢新政.血管内皮功能障碍与高血压[J].心血管病学进展,2009；30(1)：62-65.
[10]杨锦茹,魏炜.高血压患者血管内皮舒张功能与动脉弹性的关系研究[J].临床超声医学杂志,2007；9(8)：474-476.
[11]谢秀梅,李志凌,李兢,等.老年原发性高血压微量白蛋白尿与血管内皮功能及颈总动脉粥样硬化[J].高血压杂志,2006；14(2)：94-96.
[12]Anderson TJ.Asseement and treatment of endothelium dysfunction in human [J]. Am Coll Cardiol,100,34(1):631-638.
[13]胡泽平,王邦宁.血管内皮功能不全与高血压研究进展[J].医学综述,2007；13(4)：255-257.
[14]周新民.血管内皮功能及检测方法[J].内科,2008；3(2)：265-268.
[15]Gale NW, Yancopoulos GD.Growth factors acting via endothelial cell-specific receptor tyrosine kinases:VEGFs, angiopoietins, and ephrins in vascular development [J].Genes Dev,1999; 13:1055-1066.
[16]Alexander JS, Elrod JW.Extracellular matrix, junctional integrity, and matrixmetallop roteinase interactions in endothelial permeability regulation[J].J Anat,2002; 200 (8):561-574.
[17]王海燕,李晓玫,赵明辉,等.肾脏病学[M].北京：人民卫生出版社,2008,第3版：774.
[18]Msmdouh Z, Chen X, Pierini ML, et al.Targeted recycling of PECAM from endothelial surface-connected compartments during diapedesis[J].Nature,2003; 421:748-753.
[19]Schenkel AR, Mamdouh Z, Chen X, et al.CD99 plays a major role in the migration of monocytes through endothelial junctions[J]. Nat Immunol,2002; 3:145-150.
[20]Vincent PA, Xiao K, Buckley KM, et al.VE-cadherin:adhesion at arm's length[J]. Am J Physiol Cell Physiol,2004; 286:C987-C997.
[21]Gotsch U, Borges E, Bosse R, et al.VE-cadherin antibody accelerates neutrophil recruitment in vivo[J].J Cell Sci,1997; 110:583-588.
[22]Perticone F, Ceravolo R, Pujia A, et al.Prognostic significance of endothelial dysfunction in hypertensive patients [J].Circulation,2001; 104 (2):191-196.
[23]Johnson FK, Johnson RA, Peyton KJ, et al.Arginase inhibition restores arteriolar endothelial function in Dahl rats with salt-induced hypertension [J].Am J Physiol Regul Integr Comp Physiol,2005; 288 (4):R1057-1062.
[24]Klingbeil AU, John S, Schneider MP, et al.Effect of AT1 receptor blockade on endothelial function in essential hypertension [J].Am J Hypertens,2003; 16 (2): 123-128.
[25]Nitenberg A, Chemla D, Antony I. Epicardial coronary artery constriction to cold pressor test is p redictive of ardiovascular events in hypertensive patients with angiographically normal coronary arteries and without othermajor coronary risk factor [J]. Atherosclerosis,2004; 173 (1):115-123.
[26]Li LJ, Yu CM, Geng SR, et al. Endothelial dysfunction in young normotensive subjects with a family history of essential hypertension [J]. Xi'an Med Univ, 2002; 14 (1):81-85.
[27]李卫萍,孙明,周宏研,等.原发性高血压病患者血管内皮功能失调与活性氧的关系及阿托伐他汀治疗的影响[J].中国现代医学杂志,2005；15(7):1096-1103.
[28]Rekhviashvili A, Tsinamdzgvrishvili B, ChkhetiaM, et al. The relationship between endothelial dysfunction and 24-hour blood pressure rhythm in patients with arterial hypertension[J]. Georgian Med News,2008; 155:13-17.
[29]Vasudevan H, Nagareddy PR, John H, et al. Gonadectomy prevents endothelial dysfunction in fructose-fed male rats, a factor contributing to the development of hypertension[J]. Am J Physiol Heart Circ Physiol,2006; 291 (6):H3058-H3064.
[30]Zizek B, Poredos P.Increased left ventricular mass and diastolic dysfunction are associated with endothelial dysfunction in normotensive offspring of subjects with essential hypertension[J]. Blood Press,2007; 16 (1):36-44.
[31]Wallace SM, Yasmin L, McEniery CM, et al. Isolated systolic hypertension is characterized by increased aortic stiffness and endothelial dysfunction [J]. Hypertension,2007; 50:228-233.
[33]Pedrinelli R, Giampietro O, Carmassi F, et al.Microalbuminuria and endothelial dysfunction in essential hypertension[J].Lancet,1994; 344 (8914):14-8.
[34]唐锦辉.一氧化氮与肾脏疾病研究进展[J].《国外医学》泌尿内科分册,1996；16(5)：216-219.
[35]Raij L.Nitric oxide and cardiovascular and renal effects[J].Osteoarthritis Cartilage, 2008; 16Suppl 2:S21-S26.
[36]Hayakawa H, Raij L. Nitric oxide synthase activity and renal injury in genetic hypertension[J]. Hypertension,1998; 31:266-270.
[37]王正荣,罗红琳,肖静,等.天麻素对动脉血管顺应性以及血流动力学的影响[J]. 生物医学工程学杂志,1994；11(3)：197-201.
[38]任世兰,于龙顺.天麻对血管阻力和耐缺氧能力的影响[J].中草药,1992；23(6)：302-304.
[39]郑占虎,董泽宏,余靖.中药现代化研究与应用(第四卷)[M].北京：学苑出版社,1998；3335-3353.
[40]宋纯清,樊懿,黄伟晖,等.钩藤中不同成分降压作用的差异[J].中草药,2000；31(10)：762-764.
[41]王盟,刘卫.钩藤总生物碱的研究进展[J].实用医药杂志,2008；25(3)：360-362.
[42]余文海.钩藤对自发性高血压大鼠内皮的影响[J].国外医学·中医中药分册,2000；22(6)：351-352.
[43]怡悦.钩藤对自发性高血压大鼠血管内皮功能的影响[J].国外医学·中医中药分册,2000；22(1)：28-29.
[44]Gao D, Tawa R, Masaki H, et al.Protective efects of baicalein against cell damage by reactive oxygen species[J].Chem Pharm Bull,1998; 46 (9):1383-1387.
[45]应如海,王明根,吕锦芳,等.黄芩水提醇沉液对家兔动脉血压及心率的影响[J].中国中医药科技,2007；14(7)：265-266.
[46]田川智惠.杜仲叶提取物的降压作用[J].国际中医中药杂志,2006；28(4)：236.
[47]刘传瑜.老年高血压头发4种元素测定及杜仲醋浸液治疗观察[J].微量元素与健康研究,1996；13(1)：14.
[48]张瑛朝,张延敏,郭代云,等.复方杜仲叶合剂对人体降压作用的实验研究[J].中成药,2001；23(6)：418.
[49]翟文俊.杜仲叶浸提物制剂对大鼠血压及心率的影响[J].陕西教育学院学报,2004：20(4)：106.
[50]杨冀风,刘光辉.栀子提取物对大鼠阻力动脉的松驰作用[J].中成药,1999；21(9)：467-469.
[51]杨权生.桑仲汤治疗原发性高血压病70例疗效观察[J].新中医,1995； (4)：24-26.
[52]Sastre M, Dewachter I, Landreth GE, et al. Nonsteroidal anti-inflammatory drugs and peroxisome proliferators-activated receptor-agonists modulate immuno stimulatd processing of amyloid precursor protein through regulation of beta-Secretase[J].Neurosci 2003; 23 (30):796-804.
[53]Straus DS, Pascual G, Li M, et al.15-deoxy-delta 12,14-prostaglandin J2 inhibits multiple steps in the NF-kappa B signaling pathway[J].Proc Natl Acad Sci USA 2000; 97 (9):4844-4849.
[54]叶立新,王继红,黄花利.桑寄生对肾性高血压大鼠血浆β-内啡肽浓度影响的量效作用[J].中国临床康复,2005；9(27)：84-85.
[55]金琦.益母草不同剂量的药理作用和应用[J].天津中医药,2003；20(5)：51.
[1]黎磊石,刘志红,谢红浪,等.中国肾脏病学[M].北京：人民军医出版社,2008,第1版：827-848.
[2]高秀林.高血压肾损害发病机制的研究进展[J].北京医学,2007；29(9)：559-561.
[3]Palmer BE.Impaired renal autoregulation:implication for the genesis of hypertension and hypertension--induced renal injury[J]. Am J Med Sci,2001; 321:388-400.
[4]Ahmed A.Elmarabky, Jeffrey.Quigley, John D.Imig, et al.TNF-a inhibition reduces renal injury in DOCA-salt hypertensive rats[J].Am J Physiol Regul Integr Comp Physiol,2008; 294:76-83.
[5]高血压的治疗.中国高血压防治指南(2005年修订版),2005：17-25.
[6]李航,邢刚.正确防治高血压肾硬化[J/CD].中华临床医师杂志(电子版),2008；2(4)：393—396.
[7]谌贻璞.ACEI与ARB肾脏保护作用研究进展[J].中国医药导刊,2007；9(2)：122-124.
[8]杨帆,戴小华,王建彬.原发性高血压早期肾损害中医证候学观察[J].安徽中医学院学报,2007；26(2)：8-11.
[9]邓宝华.高血压肾损害早中期的中医证治体会[J].光明中医,2009；24(1)：111-112.
[10]陈政光.清热降压合剂治疗中老年高血压病及逆转早期肾损害的研究[D].2004届山东中医药大学博士学位论文.
[11]李运伦.黄连清热合剂治疗高血压病早期肾损害35例临床观察[J].新中医,2004；36(3)： 16-18.
[12]戴小华,郑林,杨帆,等.养肝益水颗粒对SHR大鼠血压及ET-1、NO的影响[J].安徽医药,2009；13(2):147-149.
[13]戴小华,郑林,杨帆,等.养肝益水颗粒对自发性高血压大鼠血压及尿MA、β2-MG的影响[J].中国中医急症,2009；18(1)：：85-86.
[14]邵正斌,杨帆,戴小华,等.养肝益水颗粒对高血压病早期肾损害尿微量蛋白及一氧化氮和内皮素的影响[J].中西医结合心脑血管病杂志,2008；6(11)：1281-1283.